临床与病理杂志2016，Vol.36Issue(4)：417-422,6.DOI:10.3978/j.issn.2095-6959.2016.04.014
氯通道对血小板活化标志物PAC-1和P-选择素的影响
The role of chloride channel on platelet activation markers PAC-1 and P-selectin
摘要
Abstract
Objective:To explore the role of chloride channel on platelet activation markers PAC-1 and P-selectin.Methods: hTe expression of ClC-3 on platelets isolated from human peripheral blood was detected by Western blotting and confocal microscopy. Using ADP (20 μM) to stimulate platelets at different points (15, 30, 60, 120, 180 min), ClC-3 protein expression was detected by Western-blot; the seven groups: the control groups, ADP (20 μM), DIDS <br> (100 μM)+ADP, NFA (100 μM)+ADP, NPPB (100 μM)+ADP, low chloride control group and low chloride +ADP (20 μM), the expressions of active platelets markers—PAC-1 and P-selectin were detected by flow cytometry.Results: The corresponding band of 85 kD could be observed by Western blot in platelets isolated from healthy adult peripheral blood; using ADP to stimulate platelets by time-dependently, ClC-3 protein expression increased signiifcantly at 15 min compared to the control group (P<0.05); the expressions of PAC-1 (32.13%±2.0%) and P-selectin (52.32%±5.31%) in ADP group were more than those in the control group (1.76%±0.41%, 1.89%±0.32%,P<0.01), while compared with the control group, the expressions of PAC-1 (8.01%±1.36%) and P-selectin (12.19%±0.84%) in low chloride control group increased; the expression levels of PAC-1 in DIDS+ADP (5.62%±1.22%), NFA+ADP (1.96%±0.54%) and NPPB +ADP (3.56%±0.79%) groups decreased comparing with ADP group (P<0.01) and the expression levels of P-selectin in DIDS+ADP (16.64%±1.52%), NFA+ADP (4.97%±0.64%) and NPPB+ADP (8.56%±2.04%) groups decreased too (P<0.01). Compared with ADP group, PAC-1 (45.26%±4.43%) and P-selectin (73.33%±3.80%) in low chloride+ADP group increased (P<0.01).Conclusion:ClC-3 chloride channel took participation in human platelet activation and chloride channel blockers could inhibit platelet activation induced by ADP, decreasing chloride level outside cells could active platelets induced by ADP.关键词
ClC-3氯通道/血小板活化/PAC-1/P-选择素Key words
ClC-3 chloride channel/platelet activation/PAC-1/P-selectin引用本文复制引用
李朋朋,黄琳燕,李玉洁,顾兵,李洪春,马萍..氯通道对血小板活化标志物PAC-1和P-选择素的影响[J].临床与病理杂志,2016,36(4):417-422,6.基金项目
国家自然科学基金青年项目(No.81402918)；江苏省科技厅青年项目(BK20140228)。hTe work was supported by the grants from the National Natural Science Foundation of China (No.81402918), the Natural Science Foundation of Jiangsu Province, China (BK20140228) （No.81402918）
